A Metabolic Switch for Cell Differentiation

Tese de doutoramento em Biologia, na especialidade de Biologia Celular, apresentada à Faculdade de Ciências e Tecnologia da Universidade de Coimbr

Mitochondrial Mechanisms of LRRK2 G2019S Penetrance

Several mutations in leucine-rich repeat kinase-2 (LRRK2) have been associated with Parkinson’s disease (PD). The most common substitution, G2019S, interferes with LRRK2 kinase activity, which is regulated by autophosphorylation. Yet, the penetrance of this gain-of-function mutation is incomplete, and thus far, few factors have been correlated with disease status in carriers. This includes (i) LRRK2 autophosphorylation in urinary exosomes, (ii) serum levels of the antioxidant urate, and (iii) abundance of mitochondrial DNA (mtDNA) transcription-associated 7S DNA. In light of a mechanistic link between LRRK2 kinase activity and mtDNA lesion formation, we previously investigated mtDNA integrity in fibroblasts from manifesting (LRRK2+/PD+) and non-manifesting carriers (LRRK2+/PD−) of the G2019S mutation as well as from aged-matched controls. In our published study, mtDNA major arc deletions correlated with PD status, with manifesting carriers presenting the highest levels. In keeping with these findings, we now further explored mitochondrial features in fibroblasts derived from LRRK2+/PD+ (n = 10), LRRK2+/PD− (n = 21), and control (n = 10) individuals. In agreement with an accumulation of mtDNA major arc deletions, we also detected reduced NADH dehydrogenase activity in the LRRK2+/PD+ group. Moreover, in affected G2019S carriers, we observed elevated mitochondrial mass and mtDNA copy numbers as well as increased expression of the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), which regulates antioxidant signaling. Taken together, these results implicate mtDNA dyshomeostasis—possibly as a consequence of impaired mitophagy—in the penetrance of LRRK2-associated PD. Our findings are a step forward in the pursuit of unveiling markers that will allow monitoring of disease progression of LRRK2 mutation carrier

Mitochondrial damage-associated inflammation highlights biomarkers in PRKN/PINK1 parkinsonism

There is increasing evidence for a role of inflammation in Parkinson’s disease. Recent research in murine models suggests that parkin and PINK1 deficiency leads to impaired mitophagy, which causes the release of mitochondrial DNA (mtDNA), thereby triggering inflammation. Specifically, the CGAS (cyclic GMP-AMP synthase)-STING (stimulator of interferon genes) pathway mitigates activation of the innate immune system, quantifiable as increased interleukin-6 (IL6) levels. However, the role of IL6 and circulating cell-free mtDNA in unaffected and affected individuals harbouring mutations in PRKN/PINK1 and idiopathic Parkinson’s disease patients remain elusive. We investigated IL6, C-reactive protein, and circulating cell-free mtDNA in serum of 245 participants in two cohorts from tertiary movement disorder centres. We performed a hypothesis-driven rank-based statistical approach adjusting for multiple testing. We detected (i) elevated IL6 levels in patients with biallelic PRKN/PINK1 mutations compared to healthy control subjects in a German cohort, supporting the concept of a role for inflammation in PRKN/PINK1-linked Parkinson’s disease. In addition, the comparison of patients with biallelic and heterozygous mutations in PRKN/PINK1 suggests a gene dosage effect. The differences in IL6 levels were validated in a second independent Italian cohort; (ii) a correlation between IL6 levels and disease duration in carriers of PRKN/PINK1 mutations, while no such association was observed for idiopathic Parkinson’s disease patients. These results highlight the potential of IL6 as progression marker in Parkinson’s disease due to PRKN/PINK1 mutations; (iii) increased circulating cell-free mtDNA serum levels in both patients with biallelic or with heterozygous PRKN/PINK1 mutations compared to idiopathic Parkinson’s disease, which is in line with previous findings in murine models. By contrast, circulating cell-free mtDNA concentrations in unaffected heterozygous carriers of PRKN/PINK1 mutations were comparable to control levels; and (iv) that circulating cell-free mtDNA levels have good predictive potential to discriminate between idiopathic Parkinson’s disease and Parkinson’s disease linked to heterozygous PRKN/PINK1 mutations, providing functional evidence for a role of heterozygous mutations in PRKN or PINK1 as Parkinson’s disease risk factor. Taken together, our study further implicates inflammation due to impaired mitophagy and subsequent mtDNA release in the pathogenesis of PRKN/PINK1-linked Parkinson’s disease. In individuals carrying mutations in PRKN/PINK1, IL6 and circulating cell-free mtDNA levels may serve as markers of Parkinson’s disease state and progression, respectively. Finally, our study suggests that targeting the immune system with anti-inflammatory medication holds the potential to influence the disease course of Parkinson’s disease, at least in this subset of patients

Observation of a new Ξb−\Xi_b^- resonance

International audienceFrom samples of pp collision data collected by the LHCb experiment at s=7, 8 and 13 TeV, corresponding to integrated luminosities of 1.0, 2.0 and 1.5  fb-1, respectively, a peak in both the Λb0K- and Ξb0π- invariant mass spectra is observed. In the quark model, radially and orbitally excited Ξb- resonances with quark content bds are expected. Referring to this peak as Ξb(6227)-, the mass and natural width are measured to be mΞb(6227)-=6226.9±2.0±0.3±0.2  MeV/c2 and ΓΞb(6227)-=18.1±5.4±1.8  MeV/c2, where the first uncertainty is statistical, the second is systematic, and the third, on mΞb(6227)-, is due to the knowledge of the Λb0 baryon mass. Relative production rates of the Ξb(6227)-→Λb0K- and Ξb(6227)-→Ξb0π- decays are also reported

Observation of the decay Bs0→D‾0K+K−B_s^0 \to \overline{D}^0 K^+ K^-

International audienceThe first observation of the Bs0→D¯0K+K- decay is reported, together with the most precise branching fraction measurement of the mode B0→D¯0K+K-. The results are obtained from an analysis of pp collision data corresponding to an integrated luminosity of 3.0  fb-1. The data were collected with the LHCb detector at center-of-mass energies of 7 and 8 TeV. The branching fraction of the B0→D¯0K+K- decay is measured relative to that of the decay B0→D¯0π+π- to be B(B0→D¯0K+K-)B(B0→D¯0π+π-)=(6.9±0.4±0.3)%, where the first uncertainty is statistical and the second is systematic. The measured branching fraction of the Bs0→D¯0K+K- decay mode relative to that of the corresponding B0 decay is B(Bs0→D¯0K+K-)B(B0→D¯0K+K-)=(93.0±8.9±6.9)%. Using the known branching fraction of B0→D¯0π+π-, the values of B(B0→D¯0K+K-)=(6.1±0.4±0.3±0.3)×10-5 and B(Bs0→D¯0K+K-)=(5.7±0.5±0.4±0.5)×10-5 are obtained, where the third uncertainties arise from the branching fraction of the decay modes B0→D¯0π+π- and B0→D¯0K+K-, respectively

Observation of Bs0→D‾∗0ϕB_s^0 \to \overline{D}^{*0} \phi and search for B0→D‾0ϕB^0 \to \overline{D}^0 \phi decays

International audienceThe first observation of the Bs0→D¯*0ϕ decay is reported, with a significance of more than seven standard deviations, from an analysis of pp collision data corresponding to an integrated luminosity of 3  fb-1, collected with the LHCb detector at center-of-mass energies of 7 and 8 TeV. The branching fraction is measured relative to that of the topologically similar decay B0→D¯0π+π- and is found to be B(Bs0→D¯*0ϕ)=(3.7±0.5±0.3±0.2)×10-5, where the first uncertainty is statistical, the second systematic, and the third from the branching fraction of the B0→D¯0π+π- decay. The fraction of longitudinal polarization in this decay is measured to be fL=(73±15±4)%. The most precise determination of the branching fraction for the Bs0→D¯0ϕ decay is also obtained, B(Bs0→D¯0ϕ)=(3.0±0.3±0.2±0.2)×10-5. An upper limit, B(B0→D¯0ϕ)<2.0 (2.3)×10-6 at 90% (95%) confidence level is set. A constraint on the ω-ϕ mixing angle δ is set at |δ|<5.2° (5.5°) at 90% (95%) confidence level

Observation of Bs0→Dˉ∗0ϕB_s^0 \to \bar{D}^{*0} \phi and search for B0→Dˉ0ϕB^0 \to \bar{D}^0 \phi decays

The first observation of the $B^0_s → \bar{D}^{*0}ϕ$ decay is reported, with a significance of more than seven standard deviations, from an analysis of pp collision data corresponding to an integrated luminosity of 3  fb$^{−1}$, collected with the LHCb detector at center-of-mass energies of 7 and 8 TeV. The branching fraction is measured relative to that of the topologically similar decay $B^0 → \bar{D}^0 π^+ π^−$ and is found to be $\mathcal{B}(B^0_s → \bar{D}^{*0}ϕ)=(3.7±0.5±0.3±0.2) × 10^{−5}$, where the first uncertainty is statistical, the second systematic, and the third from the branching fraction of the $B^0 → \bar{D}^0 π^+ π^−$ decay. The fraction of longitudinal polarization in this decay is measured to be $f_L = (73±15±4)\%$. The most precise determination of the branching fraction for the $B^0_s → \bar{D}^{0}ϕ$ decay is also obtained, $\mathcal{B}(B^0_s → \bar{D}^0 ϕ)=(3.0±0.3±0.2±0.2) × 10^{−5}$. An upper limit,$\mathcal{B}(B^0 → \bar{D}^0 ϕ) < 2.0 (2.3) × 10^{−6}$ at 90% (95%) confidence level is set. A constraint on the $ω−ϕ$ mixing angle $δ$ is set at $|δ| < 5.2°$ (5.5°) at 90% (95%) confidence level

Measurement of Angular and CP Asymmetries in D0→π+π−μ+μ−D^0\to\pi^+\pi^-\mu^+\mu^- and D0→K+K−μ+μ−D^0\to K^+K^-\mu^+\mu^- decays

International audienceThe first measurements of the forward-backward asymmetry of the dimuon pair (AFB), the triple-product asymmetry (A2ϕ), and the charge-parity-conjugation asymmetry (ACP), in D0→π+π-μ+μ- and D0→K+K-μ+μ- decays are reported. They are performed using data from proton-proton collisions collected with the LHCb experiment from 2011 to 2016, corresponding to a total integrated luminosity of 5  fb-1. The asymmetries are measured to be AFB(D0→π+π-μ+μ-)=(3.3±3.7±0.6)%, A2ϕ(D0→π+π-μ+μ-)=(-0.6±3.7±0.6)%, ACP(D0→π+π-μ+μ-)=(4.9±3.8±0.7)%, AFB(D0→K+K-μ+μ-)=(0±11±2)%, A2ϕ(D0→K+K-μ+μ-)=(9±11±1)%, ACP(D0→K+K-μ+μ-)=(0±11±2)%, where the first uncertainty is statistical and the second systematic. The asymmetries are also measured as a function of the dimuon invariant mass. The results are consistent with the standard model predictions